中文摘要：

为获得稳定来源并且具有生物学活性的重组金针菇免疫调节蛋白（Fip-fve），将fip-fve基因转至毕赤酵母GSll5中进行诱导型和组成型表达。用PCR方法从金针菇子实体基因组DNA中扩增fip-fve基因，连接至pPIC9构建诱导型表达载体pPIC9-FIP-fve，从毕赤酵母基因组DNA中扩增三磷酸甘油醛脱氢酶启动子（pgap），替换pPIC9-FIP—fve上乙醇氧化酶启动子（paoxl）构建组成型表达载体pPIC9-PGAP-FIP-fve。将线性化的两种表达载体用PEG法转化毕赤酵母GSll5，经组氨酸缺失培养基筛选和酵母菌落PCR鉴定后进行表达。结果表明，重组Fip-fve在以甲醇（1％，V/V）功为碳源进行诱导型表达4d达到最高，粗蛋白表达量为158．2mg／L，在以葡萄糖（10％）和甘油（1％，V/V）为碳源进行组成型分别在表达第4天和第5天达到最高，粗蛋白分别为46．3mg／L和29．5mg／L。SDS-PAGE及Western blotting证明重组Fip-fve已正确表达，血细胞凝集活性检测初步证明重组Fip-fve具有良好生物学活性。

英文摘要：

We transformed thefip-fve gene into Pichia pastoris GS115 for inducible and constitutive expression to obtain feasible bioactvie recombinant Fip-fve. The fip-fve gene was cloned from Flammulina velutipes fluting body by PCR and ligated to pPIC9 to construct inducible expression vector pPIC9-FIP-fve, and promotor pgap was used to replace the paoxl to construct constitutive expression vector pPIC9-PGAP-FIP-fve. These two vectors were used to transform P. pastoris by PEG method. Thefip-fve was expressed after histamine-absence screening and yeast colony PCR. The inducible expression level reached 158.2 mg/L at the fourth day and the constitutive expression level was 46.3 mg/L and 29.5 mg/L using glucose and glycerol, respectively. The SDS-PAGE and Western blotting both proved the correctness of rFip-fve, and the hemagglutination test indicats the rFip-fve＇s bioactivity.